title: Evaluation of genetic fidelity among micropropagated plants of Gloriosa superba L. using DNA-based markers--a potential medicinal plant.
abstract: Malabar glory lily (Gloriosa superba L.) is a medicinally potent plant species used for the production of alkaloid colchicine. With ever increasing demand, there is a pressing need to conserve it through biotechnological approaches. A large number of complete plantlets were obtained by direct regeneration from the non-dormant tuber explants on Murashige and Skoog (MS) medium supplemented with 2.0 mg/l 6-benzylaminopurine (BAP)+0.5 mg/l α-naphthalene acetic acid (NAA). Large number of plants can be produced in vitro under aseptic conditions, but there is always a danger of producing somaclonal variants by tissue culture technology. Thus, the genetic stability of micropropagated clones was evaluated using random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) analysis. During the study a total of 80 (50 RAPD and 30 ISSR) primers were screened, out of which 10 RAPD and 7 ISSR primers produced a total of 98 (49 RAPD and 49 ISSR) clear, distinct and reproducible amplicons. The amplification products of the regenerated plants showed similar banding patterns to that of the mother plant thus demonstrating the homogeneity of the micropropagated plants. This is the first report that evaluates the use of genetic markers to establish genetic fidelity of micropropagated G. superba using RAPD and ISSR, which can be successfully applied for the mass multiplication, germplasm conservation and further genetic transformation assays for colchicine production to meet the ever increasing demand of this medicinally potent plant for industrial and pharmaceutical uses.
authors.affiliation: Plant Tissue Culture Laboratory, Department of Botany, Kurukshetra University, Haryana 136119, India.
authors.forename: Kuldeep
authors.initials: K
authors.lastname: Yadav
authors.forename: Ashok
authors.initials: A
authors.lastname: Aggarwal
authors.forename: Narender
authors.initials: N
authors.lastname: Singh
mesh_terms: D002999:Clone Cells; D003078:Colchicine / Q000235:genetics*; D017931:DNA Primers; D018744:DNA, Plant / Q000032:analysis*; D005819:Genetic Markers; D018745:Genome, Plant; D019668:Liliaceae / Q000737:chemistry / Q000235:genetics*; D018895:Microsatellite Repeats; D010936:Plant Extracts; D018520:Plant Shoots; D058439:Plant Somatic Embryogenesis Techniques; D035281:Plant Tubers; D010946:Plants, Medicinal / Q000737:chemistry / Q000235:genetics; D019105:Random Amplified Polymorphic DNA Technique
chemical_list: D017931:DNA Primers; D018744:DNA, Plant; D005819:Genetic Markers; D010936:Plant Extracts; D003078:Colchicine
keywords: 6-benzylaminopurine; BAP; Genetic fidelity; Gloriosa superba; ISSR; Micropropagation; Molecular markers; NAA; PCR; RAPD; dNTPs; deoxyribonucleotide triphosphates; inter simple sequence repeats; polymerase chain reaction; random amplified polymorphic DNA; α-naphthalene acetic acid